Abstract
Nicotiana alata has a style-specific hydroxyproline-rich glycoprotein (the 120 kDa glycoprotein) which has properties of both extensins and AGPs [19, 20]. The 120 kDa glycoprotein is a soluble component in the extracellular matrix of the transmitting tract of styles where it accounts for ca. 9% of the total buffer-soluble protein. Here we describe the molecular cloning of a cDNA representing the gene NaPRP5 which encodes the backbone of the 120 kDa glycoprotein. Expression of mRNA is restricted to styles, consistent with observations on the distribution of the 120 kDa glycoprotein. Levels of accumulation of the transcript encoding the 120 kDa protein backbone are not altered significantly by pollination with either compatible or incompatible pollen. The protein backbone of the 120 kDa glycoprotein, as predicted by the cDNA sequence, is composed of three distinct domains. The sequence of these domains, together with linkage analysis of the carbohydrate component of the 120 kDa glycoprotein, allows predictions of the likely distribution of substituent glycosyl chains along the protein backbone. The similarity of the C-terminal domains of the 120 kDa glycoprotein and GaRSGP, the galactose-rich style glycoprotein of N. alata, is consistent with the two molecules sharing a common antigenic domain in their backbones [31]. The sharing of domains between distinct hydroxyproline-rich glycoproteins suggests that identification of a glycoprotein of this class solely by its protein or carbohydrate epitope is not valid.
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References
Chen C-G, Cornish EC, Clarke AE: Specific expression of an extensin-like gene in the style of Nicotiana alata. Plant Cell 4: 1053–1062 (1992).
Chen C-G, Mau S-L, Clarke AE: Nucleotide sequence and style-specific expression of a novel proline-rich protein gene from Nicotiana alata. Plant Mol Biol 21: 391–395 (1993).
Chen C-G, Pu Z-Y, Moritz RL, Simpson RJ, Bacic A, Clarke AE, Mau S-L: Molecular cloning of a gene encoding an arabinogalactan-protein from pear (Pyrus communis) cell suspension culture. Proc Natl Acad Sci USA 91: 10305–10309 (1994).
Chen J, Varner JE: Isolation and characterization of cDNA clones for carrot extensin and a proline-rich 33-kDa protein. Proc Natl Acad Sci USA 82: 4399–4403 (1985).
Cheung AY, May B, Kawata EE, Gu Q, Wu H-W: Characterization of cDNAs for stylar transmitting tissue-specific prolinerich proteins in tobacco. Plant J 3: 151–160 (1993).
Cotton JLS, Ross CW, Byrne DH, Colbert JT: Down-regulation of phytochorme mRNA abundance by red light and benzyladenine in etiolated cumcumber cotyledons. Plant Mol Biol 14: 707–719 (1990).
Du H, Clarke AE, Bacic A: Arabinogalactan-proteins: a class of extracellular matrix proteoglycans involved in plant growth and development.Trends Cell Biol 6: 411–414 (1996).
Du H, Simpson RJ, Clarke AE, Bacic A: Molecular characterization of a stigma-specific gene encoding an arabinogalactanprotein (AGP) from Nicotiana alata.Plant J9: 313–323 (1996).
Du H, Simpson RJ, Moritz RL, Clarke AE, Bacic A: Isolation of the protein backbone of an arabinogalactan-protein from the styles of Nicotiana alata and characterization of a corresponding cDNA.Plant Cell 6: 1643–1653 (1994).
Fincher GB, Stone BA, Clarke AE: Arabinogalactan-proteins: structure, biosynthesis and function. Annu Rev Plant Physiol 34: 47–70 (1983).
Gleeson PA, McNamara M, Wettenhall REH, Stone BA, Fincher GB: Characterization of the hydroxyproline-rich protein core of an arabinogalactan-protein secreted from suspensioncultured Loliummultiflorum (Italian ryegrass) endosperm cells. Biochem J 264: 857–862 (1989).
Goldman MHS, Pezzotti M, Seurinck J, Mariani C: Developmental expression of tobacco pistil-specific genes encoding novel extensin-like proteins. Plant Cell 4: 1041–1051 (1992).
Hay B, Short JM: ExassistTM helper phage and SOLRTMcells for Lambda ZAP II excisions. Strategies 5: 16–18 (1992).
Heslop-Harrison J: Pollen germination and pollen-tube growth. Int Rev Cytol 107: 1–78 (1987).
Kieliszewski MJ, Lamport DTA: Extensin: repetitive motifs, functional sites, post-translational codes, and phylogeny. Plant J 5: 157–172 (1994).
Labarca C, Loewus F: The nutritional role of pistil exudate in pollen tube wall formation in Lilium longiflorum. I. Utilization of injected stigmatic exudate. Plant Physiol 50: 7–14 (1972).
Labarca C, Loewus F: The nutritional role of pistil exudate in pollen tube wall formation in Lilium longiflorum. II Production and utilization of exudate from stigma and stylar canal. Plant Physiol 52: 87–92 (1973).
Landon M: Cleavage at aspartyl-prolyl bonds. Meth Enzymol47: 145–149 (1977).
Lind JL: Characterization of an hydroxyproline-rich glycoprotein from the style of Nicotiana alata. Ph.D thesis, University of Melbourne (1993).
Lind JL, Bacic A, Clarke AE, Anderson MA: A stylespecific hydroxyproline-rich glycoprotein with properties of both extensins and arabinogalactan proteins. Plant J 6: 491– 502 (1994).
Lind JL, Bönig I, Clarke AE, Anderson MA: A style-specific 120-kDa glycoprotein enters pollen tubes of Nicotiana alata in vivo.Sex Plant Reprod 9: 75–86 (1996).
Marchuk D, Drumm M, Saulino A, Collins FS: Constuction of T-vectors, a rapid and general system for direct cloning of unmodified PCR products. Nucl Acids Res 19: 1154 (1991).
Masuda Y, Nirasawa S, Nakaya K, Kurihara Y: Cloning and sequencing of a cDNA clone encoding a taste-modifying protein, miraculin. Gene 161: 175–177 (1995).
Myerson D: Producing single-stranded DNA probes with Taq DNA polymerase: a high yield protocol. Biotechniques 10: 35–38 (1991).
Nielsen H, Engelbrecht J, Brunak S, Heijne Gv: Identification of prokaryotic and eukaryotic signal peptides and prediction of their cleavage sites. Prot Enging 10: 1–6 (1997).
Ogretmen B, Ratajczak H, Kats A, Stark BC, Gendel SM: Effects of staining of RNA with ethidium bromide before electrophoresis on performance of northern blots. Biotechniques 14: 932–935 (1993).
Rubinstein AL, Broadwater AH, Lowrey KB, Bedinger PA: Pex1, a pollen-specific gene with an extensin-like domain. Proc Natl Acad Sci USA 92: 3086–3090 (1995).
Sambrook F, Fritsch EF, Maniatus T: Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1989).
Showalter AM, Varner JE: Plant hydroxyproline-rich glycoproteins.Biochem Plants 15: 485–520 (1989).
Simpson RJ, Ward LD, Reid GE, Batterham MP, Moritz RL: Peptide mapping and internal sequencing of proteins electroblotted from two-dimenional gels onto polyvinylidene difluoride membranes.J Chromotogr476: 345–361 (1989).
Sommer-Knudsen J, Clarke AE, Bacic A: A galactose-rich, cell-wall glycoprotein from styles of Nicotiana alata.Plant J 9:71–83 (1996).
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Schultz, C.J., Hauser, K., Lind, J.L. et al. Molecular characterisation of a cDNA sequence encoding the backbone of a style-specific 120 kDa glycoprotein which has features of both extensins and arabinogalactan proteins. Plant Mol Biol 35, 833–845 (1997). https://doi.org/10.1023/A:1005816520060
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DOI: https://doi.org/10.1023/A:1005816520060